Effect of polyamine depletion on DNA damage and repair following UV irradiation of HeLa cells.
Treatment of HeLa cells with the polyamine biosynthesis inhibitors, methylglyoxal bis(guanylhydrazone) (MGBG), difluoromethylornithine (DFMO) or a combination of the two, resulted in reduction in cellular polyamine levels. Analysis of UV light-induced DNA damage and repair in these polyamine depleted cells revealed distinct differences in the repair process relative to that seen in cells possessing a normal polyamine complement. Initial yield of thymine dimers and rate of removal of these lesions from cellular DNA appeared normal in polyamine-depleted cells. However, depleted cells exhibited retarded sealing of DNA strand breaks resulting from cellular repair processes, reduced repair synthesis and an increased sensitivity to UV killing. Incision at damaged sites was not affected since ara-C repair-dependent breaks accumulated in a normal fashion. Molecular analysis of inhibited repair sites by exonuclease III and T4 DNA ligase probes suggest that the strand interruptions consist of gaps rather than ligatable nicks, consistent with an interpretation of the repair defect being at the gap-filling stage rather than the ligation step. Observed patterns of differential polyamine depletion by DFMO and MGBG, and partial reversal of repair inhibition by polyamine supplementation, suggests that polyamine depletion per se, rather than some secondary effect of inhibitor treatment, is responsible for the inhibition of repair.
Snyder
R D
RD
Merrell Dow Research Institute, Cincinnati, OH 45215.
Sunkara
P S
PS
D003114:Colony-Forming Units Assay; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication / Q000187:drug effects / Q000528:radiation effects; D004273:DNA, Neoplasm / Q000528:radiation effects*; D000518:Eflornithine / Q000494:pharmacology*; D006367:HeLa Cells / Q000378:metabolism / Q000528:radiation effects; D006801:Humans; D008935:Mitoguazone / Q000494:pharmacology*; D011073:Polyamines / Q000378:metabolism* / Q000494:pharmacology; D014466:Ultraviolet Rays
D004273:DNA, Neoplasm; D011073:Polyamines; D008935:Mitoguazone; D000518:Eflornithine